title: Rapid, Facile Detection of Heterodimer Partners for Target Human G-Protein-Coupled Receptors Using a Modified Split-Ubiquitin Membrane Yeast Two-Hybrid System.
abstract: Potentially immeasurable heterodimer combinations of human G-protein-coupled receptors (GPCRs) result in a great deal of physiological diversity and provide a new opportunity for drug discovery. However, due to the existence of numerous combinations, the sets of GPCR dimers are almost entirely unknown and thus their dominant roles are still poorly understood. Thus, the identification of GPCR dimer pairs has been a major challenge. Here, we established a specialized method to screen potential heterodimer partners of human GPCRs based on the split-ubiquitin membrane yeast two-hybrid system. We demonstrate that the mitogen-activated protein kinase (MAPK) signal-independent method can detect ligand-induced conformational changes and rapidly identify heterodimer partners for target GPCRs. Our data present the abilities to apply for the intermolecular mapping of interactions among GPCRs and to uncover potential GPCR targets for the development of new therapeutic agents.
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Jun
authors.initials: J
authors.lastname: Ishii
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
mesh_terms: D019281:Dimerization; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005838:Genotype; D006801:Humans; D008024:Ligands; D044140:Receptor, Angiotensin, Type 1 / Q000737:chemistry / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000378:metabolism*; D017481:Receptors, Somatostatin / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques; D025801:Ubiquitin / Q000378:metabolism*; D015003:Yeasts / Q000235:genetics / Q000378:metabolism*
chemical_list: C544065:AGTR1 protein, human; D005656:Fungal Proteins; D008024:Ligands; D044140:Receptor, Angiotensin, Type 1; D043562:Receptors, G-Protein-Coupled; D017481:Receptors, Somatostatin; D025801:Ubiquitin; C094573:somatostatin receptor 5